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Resistant starch (RS) confers many health benefits, mostly through the microbial production of SCFAs, but foods containing appreciable RS are limited. High-amylose wheat (HAW) is high in RS and lowers the glycemic response of foods, but whether it can improve gastrointestinal health measures is unknown.The objective of this study was to determine whether daily consumption of HAW food products improved markers of gastrointestinal health in healthy men and women compared with similar foods made from conventional wheat.Eighty healthy adults (47 women and 33 men) were enrolled in a 4-arm parallel, randomized-controlled, double-blind trial. After a 2-wk low-dietary fiber run-in period, they were randomly allocated to 1 of 4 treatment groups: low-amylose wheat (LAW)-refined (LAW-R), LAW-wholemeal (LAW-W), HAW-refined (HAW-R), and HAW-wholemeal (HAW-W) and consumed the assigned test bread (160 g/d) and biscuits (75 g/d) for 4 wk. Fecal biochemical markers were measured at baseline and 4 wk. Microbial abundance and diversity were quantified using 16S ribosomal RNA sequencing and perceived gut comfort by a semiquantitative questionnaire completed at baseline, 2 wk, and 4 wk.HAW showed similar effects on fecal output and excretion of total SCFA compared with LAW, but changes were observed in secondary measures for the refined treatment groups. At 4 wk, the HAW-R group had 38% higher fecal butyrate excretion than the LAW-R group (P 0.05), and higher fecal SCFA-producing bacteria, Roseburia inulinivorans (P 0.001), than at baseline. In comparison with baseline, LAW-R increased fecal p-cresol concentration, and fecal abundance of a p-cresol-producing bacterium, Clostridium from the Peptostreptococcaceae family, but both were reduced by HAW-R. Amylose level did not affect measures of fecal consistency or adversely affecting digestive comfort.Increasing RS intake of healthy adults by substituting refined conventional wheat with refined HAW modulates fecal metabolites and microbes associated with gastrointestinal health.This trial was registered at anzctr.org.au as ACTRN12618001060235.

BACKGROUND Conventional wheat-based foods contain high concentrations of readily digestible starch that commonly give these foods a high postprandial glycemic response and may contribute to the development of type 2 diabetes and cardiovascular disease. OBJECTIVES The aim of this study was to determine if bread made from high-amylose wheat (HAW) and enriched in resistant starch dampens postprandial glycemia compared with bread made from conventional low-amylose wheat (LAW). METHODS This single-center, randomized, double-blinded, crossover controlled study involved 7 consecutive weekly visits. On separate mornings, 20 healthy nondiabetic men and women (mean age 30 ± 3 y; body mass index 23 ± 0.7 kg/m2) consumed a glucose beverage or 4 different breads (each 121 g); LAW-R (refined), LAW-W (wholemeal), HAW-R, or HAW-W. The starch contents of the LAW and HAW breads were 24% and 74% amylose, respectively. Venous blood samples were collected at regular intervals before and for 3 h after the breakfast meal to measure plasma glucose, insulin, ghrelin, and incretin hormone concentrations, and the incremental area under the curve (AUC) was calculated (mmol/L × 3 h). Satiety and cravings were also measured at 30-min intervals during the postprandial period. RESULTS HAW breads had a glycemic response (AUC) that was 39% less than that achieved with conventional wheat breads (HAW 39 ± 5 mmol/L × 3 h; LAW 64 ± 5 mmol/L × 3 h; P

Background The Paleolithic diet is promoted worldwide for improved gut health. However, there is little evidence available to support these claims, with existing literature examining anthropometric and cardiometabolic outcomes. Objective To determine the association between dietary intake, markers of colonic health, microbiota, and serum trimethylamine-N-oxide (TMAO), a gut-derived metabolite associated with cardiovascular disease. Design In a cross-sectional design, long-term (n = 44, > 1 year) self-reported followers of a Paleolithic diet (PD) and controls (n = 47) consuming a diet typical of national recommendations were recruited. Diets were assessed via 3-day weighed diet records; 48-h stool for short chain fatty acids using GC/MS, microbial composition via 16S rRNA sequencing of the V4 region using Illumina MiSeq. TMAO was quantified using LC–MS/MS. Results Participants were grouped according to PD adherence; namely excluding grains and dairy products. Strict Paleolithic (SP) (n = 22) and Pseudo-Paleolithic (PP) (n = 22) groups were formed. General linear modelling with age, gender, energy intake and body fat percentage as covariates assessed differences between groups. Intake of resistant starch was lower in both Paleolithic groups, compared to controls [2.62, 1.26 vs 4.48 g/day (P P Hungatella in both Paleolithic groups (P P r = − 0.34, P Conclusions Although the PD is promoted for improved gut health, results indicate long-term adherence is associated with different gut microbiota and increased TMAO. A variety of fiber components, including whole grain sources may be required to maintain gut and cardiovascular health. Clinical trial registrations Australian and New Zealand Clinical Trial Registry (ANZCTRN12616001703493).

Summary Reliably generating rice varieties with low glycaemic index (GI) is an important nutritional intervention given the high rates of Type II diabetes incidences in Asia where rice is staple diet. We integrated a genome‐wide association study (GWAS) with a transcriptome‐wide association study (TWAS) to determine the genetic basis of the GI in rice. GWAS utilized 305 re‐sequenced diverse indica panel comprising ~2.4 million single nucleotide polymorphisms (SNPs) enriched in genic regions. A novel association signal was detected at a synonymous SNP in exon 2 of LOC_Os05g03600 for intermediate‐to‐high GI phenotypic variation. Another major hotspot region was predicted for contributing intermediate‐to‐high GI variation, involves 26 genes on chromosome 6 (GI6.1). These set of genes included GBSSI, two hydrolase genes, genes involved in signalling and chromatin modification. The TWAS and methylome sequencing data revealed cis‐acting functionally relevant genetic variants with differential methylation patterns in the hot spot GI6.1 region, narrowing the target to 13 genes. Conversely, the promoter region of GBSSI and its alternative splicing allele (G allele of Wx a) explained the intermediate‐to‐high GI variation. A SNP (C˃T) at exon‐10 was also highlighted in the preceding analyses to influence final viscosity (FV), which is independent of amylose content/GI . The low GI line with GC haplotype confirmed soft texture, while other two low GI lines with GT haplotype were characterized as hard and cohesive. The low GI lines were further confirmed through clinical in vivo studies. Gene regulatory network analysis highlighted the role of the non‐starch polysaccharide pathway in lowering GI.

Background Diet-related non communicable diseases (NCDs) impose a heavy burden on human health worldwide. Rice is a good target for diet-related disease prevention strategies because it is widely consumed. Liu et al. demonstrated that increasing the number of cell layers and thickness of putative aleurone in ta2-1 (thick aleurone 2-1) mutant rice enhances simultaneously the content of multiple micronutrients. However, the increases of aleurone-associated nutrients were not proportional to the increases in the aleurone thickness. Results In this study, first, cytohistological analyses and transmission electron microscopy demonstrated that the multilayer in ta2-1 exhibited aleurone cell structural features. Second, we detected an increase in insoluble fibre and insoluble bound-phenolic compounds, a shift in aleurone-specific NNSP (neutral non-starch polysaccharide) profile, enhancement of phytate and minerals such as iron, zinc, potassium, magnesium, sulphur, and manganese, enrichment of triacylglycerol and phosphatidylcholine but slight reduction in free fatty acid, and an increase in oleic fatty acid composition. Conclusion These findings support our hypothesis that the expanded aleurone-like layers in ta2-1 maintained the distinctive aleurone features and composition associated with true aleurone. We provide perspectives to achieve even greater filling of this expanded micronutrient sink to alleviate the burden of NCDs.

The human ileostomy model, widely considered the benchmark for determining in vivo starch digestibility, has disadvantages. The ileorectostomised rat model (IRM) is a possible surrogate but evidence as to its validity is scant. In this preliminary study, the resistant starch (RS) content of test breads made from refined low (LAW-R) and high amylose wheat (HAW-R) flours was established in a randomised cross-over trial involving six human ileostomy participants. Starch digestibility of refined breads and diets made from these flours was then evaluated in ileorectostomised rats using a similar experimental format. Physical performance measures and other data were also collected for the rat model. The amount of RS in the low- and high-amylose breads as measured using the human model was 0.8 &plusmn, 0.1 and 6.5 &plusmn, 0.3 g/100 g, respectively. The RS level of HAW-R bread determined using ileorectostomised rats was 5.5 &plusmn, 0.8 g/100 g, about 15% less than that recorded in the human study, whereas for conventional wheat breads the models produced similar RS values. While offering promise, further validation using a wide variety of starchy food products is needed before the IRM can be considered an acceptable alternative for RS determination.

The enzyme starch synthase IIa (SSIIa) in cereals has catalytic and regulatory roles during the synthesis of amylopectin that influences the functional properties of the grain. Rice endosperm SSIIa is more active in indica accessions compared to japonica lines due to functional SNP variations in the coding region of the structural gene. In this study, downregulating the expression of japonica-type SSIIa in Nipponbare endosperm resulted in either shrunken or opaque grains with an elevated proportion of A-type starch granules. Shrunken seeds had severely reduced starch content and could not be maintained in succeeding generations. In comparison, the opaque grain morphology was the result of weaker down-regulation of SSIIa which led to an elevated proportion of short-chain amylopectin (DP 6-12) and a concomitant reduction in the proportion of medium-chain amylopectin (DP 13-36). The peak gelatinization temperature of starch and the estimated glycemic score of cooked grain as measured by the starch hydrolysis index were significantly reduced. These results highlight the important role of medium-chain amylopectin in influencing the functional properties of rice grains, including its digestibility. The structural, regulatory and nutritional implications of down-regulated japonica-type SSIIa in rice endosperm are discussed.

The Paleolithic diet excludes two major sources of fibre, grains and legumes. However, it is not known whether this results in changes to resistant starch (RS) consumption. Serum trimethylamine-N-oxide (TMAO) is produced mainly from colonic fermentation and hepatic conversion of animal protein and is implicated in CVD, but changes in RS intake may alter concentrations. We aimed to determine whether intake of RS and serum concentrations of TMAO varied in response to either the Paleolithic or the Australian Guide to Healthy Eating (AGHE) diets and whether this was related to changes in food group consumption. A total of thirty-nine women (mean age 47 (sd 13) years, BMI 27 (sd 4) kg/m2) were randomised to AGHE (n 17) or Paleolithic diets (n 22) for 4 weeks. Serum TMAO concentrations were measured using liquid chromatography–MS; food groups, fibre and RS intake were estimated from weighed food records. The change in TMAO concentrations between groups (Paleolithic 3·39 μmv. AGHE 1·19 μm, P = 0·654) did not reach significance despite greater red meat and egg consumption in the Paleolithic group (0·65 serves/d; 95 % CI 0·2, 1·1; P P P

Non-communicable diseases such as diabetes, cardiovascular diseases and certain cancers are now the leading cause of death and disability, and their prevalence is rising worldwide. Poor diet is a major modifiable risk factor but changing eating habits has had limited success. Enhancing the nutritional quality of staple foods offers a complimentary intervention strategy for alleviating the burden of diet-related chronic disease. Wholegrain cereals, such as wheat, are prime targets. Their nutritional credentials and health-promoting potential are well established. Wheat is the major source of protein, minerals and vitamins, and dietary fibre for most people. Importantly, wheat is a versatile ingredient for producing foods that have high consumer appeal. Their popularity is expanding globally and small improvements in wheat grain composition conceivably translate to substantial dietary change across entire populations. A newly developed, high amylose wheat line illustrates the capacity for developing healthier processed staple food products with potential for remediating chronic disease risk. Dietary fibre density of this novel grain is markedly improved due to a >10 fold increase in resistant starch content. The sensory attributes of finished products are not compromised and preliminary studies demonstrate that consumption of the new wheat significantly improves indices of metabolic and digestive health.

In adults, fermentation of high amylose maize starch (HAMS), a resistant starch (RS), has a prebiotic effect. Were such a capacity to exist in infants, intake of RS might programme the gut microbiota during a critical developmental period. This study aimed to determine if infant faecal inocula possess the capacity to ferment HAMS or acetylated-HAMS (HAMSA) and characterise associated changes to microbial composition. Faecal samples were collected from 17 healthy infants at two timepoints: Preweaning and within 10 weeks of first solids. Fermentation was assessed using in vitro batch fermentation. Following 24 h incubation, pH, short-chain fatty acid (SCFA) production and microbial composition were compared to parallel control incubations. In preweaning infants, there was a significant decrease at 24 h in pH between control and HAMS incubations and a significant increase in the production of total SCFAs, indicating fermentation. Fermentation of HAMS increased further following commencement of solids. Fermentation of RS with weaning faecal inocula increased Shannon&rsquo, s diversity index (H) and was associated with increased abundance of Bifidobacterium and Bacteroides. In conclusion, the faecal inocula from infants is capable of RS fermentation, independent of stage of weaning, but introduction of solids increases this fermentation capacity. RS may thus function as a novel infant prebiotic.

Long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) may be more bioavailable from krill oil compared to fish oil due to their phospholipid structure. We tested whether a microencapsulated krill and tuna oil blend (ME-TOKO) provided greater LC n-3 PUFA bioavailability, improved blood lipid profiles and increased intestinal contractility compared to microencapsulated tuna oil (ME-TO). Rats were divided into three groups to receive isocaloric diets containing ME-TO, ME-TOKO and microencapsulated olive oil (ME-OO) at 0.3 or 2 g/100 g for 4 weeks. Final body and organ weights, feed intake and waste output were similar. ME-TOKO rats had higher plasma total LC n-3 PUFA levels compared to ME-TO, but liver LC n-3 PUFA levels and plasma triglyceride and cholesterol levels were similar in non-fasted rats. Diets containing 2% ME-TO and ME-TOKO also showed similar increases in ileal contractility. In summary, ME-TO bioavailability of LC n-3 PUFA was similar to ME-TOKO.

Using barley cultivars differing widely in β-glucan content, we aimed to determine their effects on small intestinal macronutrient digestion in 24 ileorectostomised rats. The rats were fed 1 of 4 experimental diets, each containing a different barley variety, for 11 d. The diets had a content of 0, 2.1, 2.6 and 4.3 g of β-glucan/100 g. Feed intake and faecal excretion of fat, protein, starch, and non-starch polysaccharides were determined in the final 5 d of the study and apparent macronutrient digestibility calculated. Higher dietary levels of β-glucan (2.6% and 4.3%) lowered feed intake (by 15 and 19%, respectively) but final body weight was only lowered by the 4.3% β-glucan diet relative to rats fed the 0% β-glucan diet (all ps

Intestinal fermentation of inulin-type fructans, including oligofructose, can modulate adiposity, improve energy regulation, and increase mineral absorption. We aimed to determine whether cereal fructans had greater effects on reducing adiposity and improving mineral absorption compared with oligofructose. Thirty-two male Sprague–Dawley rats were randomly assigned to one of four dietary treatments that contained 0% fructan (control), or 5% fructan provided by oligofructose (OF), a barley grain fraction (BGF), or a wheat stem fraction (WSF). After 1 week on the diets, mineral absorption and retention was assessed. At 4 weeks, blood samples were collected for gut hormone analysis, adipose depots were removed and weighed, and caecal digesta was analyzed for pH and short-chain fatty acids (SCFA). The BGF and WSF, but not OF, had lower total visceral fat weights than the Control (p

A possible causal role of the gut microbiota in human obesity is capturing interest. Recent experimental evidence and mechanistic hypotheses suggest that a ‘dysbiotic’ large bowel microbiota, induced mainly by poor diet, increases dietary energy bioavailability and storage in the host. However, research findings in both animals and humans are inconsistent and whether an altered gut microbiota meaningfully impacts host energetics remains an open question. Future intervention studies must control diet and other lifestyle factors that profoundly influence the composition and activity of the intestinal microbiota to define its potential role in and contribution to the human obesity problem.

Objectives: The aim of this study was to investigate how a moderate increase in dietary meat content combined (or not) with soluble fibre would influence protein digestion as well as digesta characteristics and flow. Methods: Four groups of pigs were fed Western-style diets (high-protein/high-fat) containing two types of barbecued red meat, one with and one without a wheat arabinoxylan-rich fraction. After 4 wk, digesta samples were collected from small and large intestinal sites and analyzed for protein, amino acids, dry matter, and acid-insoluble ash. Tissue samples were also collected from each site. Results: Arabinoxylan consumption led to somewhat lower apparent protein digestibility within the small and large intestines as well as shorter mean retention times. This suggests that the lowered protein digestibility is due, at least partly, to shorter access time to digestive proteases and absorptive surfaces. Additionally, digesta mass was higher in pigs fed arabinoxylan while dry matter (%) was lower, indicating an increased digesta water-holding capacity due to the presence of a soluble dietary fiber. Conclusion: Data showed that solubilized wheat arabinoxylan provides potential health benefits through decreased protein digestibility, increased digesta mass, and reduced mean retention time, even for diets with a moderately higher protein content. These factors are associated with efficiency of digestion and satiety, both of which have implications for prevention of obesity and other health disorders.

Summary Resistant starch (RS), a type of dietary fibre, plays an important role in human health; however, the content of RS in most modern processed starchy foods is low. Cereal starch, when structurally manipulated through a modified starch biosynthetic pathway to greatly increase the amylose content, could be an important food source of RS. Transgenic studies have previously revealed the requirement of simultaneous down-regulation of two starch branching enzyme (SBE) II isoforms both located on the long arm of chromosome 2, namely SBEIIa and SBEIIb, to elevate the amylose content in wheat from ~25% to ~75%. The current study revealed close proximity of genes encoding SBEIIa and SBEIIb isoforms in wheat with a genetic distance of 0.5 cM on chromosome 2B. A series of deletion and single nucleotide polymorphism (SNP) loss of function alleles in SBEIIa, SBEIIb or both was isolated from two different wheat populations. A breeding strategy to combine deletions and SNPs generated wheat genotypes with altered expression levels of SBEIIa and SBEIIb, elevating the amylose content to an unprecedented ~85%, with a marked concomitant increase in RS content. Biochemical assays were used to confirm the complete absence in the grain of expression of SBEIIa from all three genomes in combination with the absence of SBEIIb from one of the genomes.

Fermentation of oat and barley β-glucans is believed to mediate in part their metabolic health benefits, but the exact mechanisms remain unclear. In this study, we sought to test the hypothesis that barley β-glucan fermentation raises circulating incretin hormone levels and improves glucose control, independent of other grain components. Male Sprague-Dawley rats (n = 30) were fed a high-fat diet for 6 weeks and then randomly allocated to 1 of 3 dietary treatments for 2 weeks. The low- (LBG, 0% β-glucan) and high- (HBG, 3% β-glucan) β-glucan diets contained 25% wholegrain barley and similar levels of insoluble dietary fiber, available carbohydrate, and energy. A low-fiber diet (basal) was included for comparison. Immediately prior to the dietary intervention, gastric emptying rate (using the (13)C-octanoic breath test) and postprandial glycemic response of each diet were determined. At the end of the study, circulating gut hormone levels were determined; and a glucose tolerance test was performed. The rats were then killed, and indices of cecal fermentation were assessed. Diet did not affect live weight; however, the HBG diet, compared to basal and LBG, reduced food intake, tended to slow gastric emptying, increased cecal digesta mass and individual and total short-chain fatty acid pools, and lowered digesta pH. In contrast, circulating levels of glucose, insulin, gastric-inhibitory peptide, and glucagon-like peptide-1, and glucose tolerance were unaffected by diet. In conclusion, wholegrain barley β-glucan suppressed feed intake and increased cecal fermentation but did not improve postprandial glucose control or insulin sensitivity.

cope A recent study revealed that the accumulation of gut microbiota-produced acetate (GMPA) led to insulin over-secretion and obesity symptom. To further develop this scientific point, the effect of resistant starch (RS) or exogenous acetate carried by RS (RSA) in the gut on metabolic syndrome is investigated using diet-induced obese rats. Methods and results The metabonomics analysis shows that the gut of rats in the RSA group generate more butyrate in both serum and feces rather than acetate compared to the rats in RS group, indicating the conversion among metabolites, in particular from acetate to butyrate via gut microbiota. Consistently, the gut microbiome uses acetate as a substrate to produce butyrate, such as Coprococcus, Faecalibacterium, Roseburia, and Eubacterium and was highly promoted in RSA group, which further supports the metabolic conversion. This is the first report to reveal the accumulation of gut microbiota-produced butyrate (GMPB) but not GMPA significantly enriched AMPK signaling pathway with reduced expression of lipogenesis-associated genes for suppressing sphingosines and ceramides biosynthesis to trigger insulin sensitivity. Conclusion Gut microbiome profile and lipogenesis pathway are regulated by GMPB, which substantially influences energy harvesting in the gut from patterns opposed to GMPA.

Rice lines with slower starch digestibility provide opportunities in mitigating the global rise in type II diabetes and related non-communicable diseases. However, screening for low glycemic index (GI) in rice breeding programs is not possible due to time and cost constraints. This study evaluated the feasibility of using in vitro cooked grain amylolysis, starch mobilization patterns during seed germination, and variation in starch structure and composition in the mature seed to differentiate patterns of starch digestibility. Mobilization patterns of total starch, resistant starch, amylose and amylopectin chains, and free sugars during seed germination revealed that the process is analogous to digestion in the human gastrointestinal tract. The combination of these biochemical markers can be used as an alternative measure to predict GI. Additionally, transcriptome analysis of stored mRNA transcripts in high and low GI lines detected differences in starch metabolism and confirmed the importance of seed storage pathways in influencing digestibility. Pathway analyses supported by metabolomics data revealed that resistant starch, cell wall non-starch polysaccharides and flavonoids potentially contribute to slower digestibility. These new insights can guide precision breeding programs to produce low GI rice with acceptable cooking quality to help mitigate the burden of diet-associated lifestyle diseases.

BACKGROUND: Rats used in nutritional studies are often kept in wire-based cages to prevent ingestion of bedding and minimise ingestion of faeces. However, wire-based cages are criticised because of potential negative animal welfare implications. This study investigated the effects of wire and solid-based cages with corncob bedding on large bowel fermentation and microbiota. Rats were group housed in wire or solid-based cages and fed either a low-fibre (LF) diet or a high-fibre (HF) diet composed of resistant starch for 4 weeks. RESULTS: Bedding material was observed in faeces of rats housed in solid-based cages. Caging type and diet altered large bowel fermentation variables and bacterial populations. Caecal digesta weight was lower in rats fed HF diet and maintained on bedding than in HF-fed rats maintained on wire. Bacteria abundance associated with fibre fermentation was higher in LF-diet fed rats maintained on bedding compared with LF-fed rats housed on wire. CONCLUSION: Maintaining rats in solid-based cages with corncob bedding alters large bowel fermentation and bacterial communities owing to ingestion of bedding. These changes may confound outcomes of nutritional studies, particularly those investigating the health effects of fibres. The use of wire-based caging may be justified in research of this type. © 2014 Society of Chemical Industry

Author version made available in accordance with the publisher's policy., High red meat intake is associated with increased colorectal cancer (CRC) risk, while resistant starch is probably protective. Resistant starch fermentation produces butyrate, which can alter microRNA (miRNA) levels in CRC cells in vitro; effects of red meat and resistant starch on miRNA expression in vivo were unknown. This study examined whether a high red meat (HRM) diet altered miRNA expression in rectal mucosa tissue of healthy volunteers, and if supplementation with butyrylated resistant starch (HRM+HAMSB) modified this response. In a randomised cross-over design, twenty-three volunteers undertook four 4-week dietary interventions; an HRM diet (300 g/day lean red meat) and an HRM+HAMSB diet (HRM with 40 g/day butyrylated high amylose maize starch), preceded by an entry diet and separated by a washout. Faecal butyrate increased with the HRM+HAMSB diet. Levels of oncogenic mature miRNAs, including miR-17-92 cluster miRNAs and miR-21, increased in the rectal mucosa with the HRM diet, while the HRM+HAMSB diet restored miR-17-92 miRNAs, but not miR-21, to baseline levels. Elevated miR-17-92 and miR-21 in the HRM diet corresponded with increased cell proliferation, and a decrease in miR-17-92 target gene transcript levels, including CDKN1A. The oncogenic miR-17-92 cluster is differentially regulated by dietary factors that increase or decrease risk for CRC, and this may explain, at least in part, the respective risk profiles of high red meat and resistant starch. These findings support increased resistant starch consumption as a means of reducing risk associated with an HRM diet.

Objective A low FODMAP (Fermentable Oligosaccharides, Disaccharides, Monosaccharides And Polyols) diet reduces symptoms of IBS, but reduction of potential prebiotic and fermentative effects might adversely affect the colonic microenvironment. The effects of a low FODMAP diet with a typical Australian diet on biomarkers of colonic health were compared in a single-blinded, randomised, cross-over trial. Design Twenty-seven IBS and six healthy subjects were randomly allocated one of two 21-day provided diets, differing only in FODMAP content (mean (95% CI) low 3.05 (1.86 to 4.25) g/day vs Australian 23.7 (16.9 to 30.6) g/day), and then crossed over to the other diet with ≥21-day washout period. Faeces passed over a 5-day run-in on their habitual diet and from day 17 to day 21 of the interventional diets were pooled, and pH, short-chain fatty acid concentrations and bacterial abundance and diversity were assessed. Results Faecal indices were similar in IBS and healthy subjects during habitual diets. The low FODMAP diet was associated with higher faecal pH (7.37 (7.23 to 7.51) vs 7.16 (7.02 to 7.30); p=0.001), similar short-chain fatty acid concentrations, greater microbial diversity and reduced total bacterial abundance (9.63 (9.53 to 9.73) vs 9.83 (9.72 to 9.93) log 10 copies/g; p Clostridium cluster XIVa (median ratio 6.62; p Akkermansia muciniphila (19.3; p Ruminococcus torques . Conclusions Diets differing in FODMAP content have marked effects on gut microbiota composition. The implications of long-term reduction of intake of FODMAPs require elucidation. Trial registration number ACTRN12612001185853.

The use of small animal models for studying postprandial changes in circulating nutrients, hormones and metabolic biomarkers is hampered by the limited quantity of blood that can be withdrawn for analysis. Here, we describe the development of an unrestrained, meal-fed rat model, having a permanent or temporary vascular cannula that permits repeated blood sampling. The applicability and performance of the model were evaluated in a series of experiments on acute glycaemic and insulinaemic responses to carbohydrate-based test meals.A test food containing 0.4 g carbohydrate raised blood glucose by 1.5 mmol LThis improved small animal model affords new opportunities to screen foods for nutrient bioavailability and explore metabolic mechanisms mediating responses to food consumption. © 2016 Society of Chemical Industry.

Scope: Soluble dietary fibres have shown to have lipid reducing properties. However, their mechanisms of action are still unclear. The present study investigated how a soluble wheat arabinoxylan-rich fraction (AXRF) fed to pigs used as a human model reduced blood triglycerides. Methods and results: After 4 weeks on the experimental diets, blood from the jugular (JV) and hepatic portal (HPV) veins, bile from the gall bladder, and digesta samples from four sites of the small intestine (SI) and cecum were collected. The results showed that the AXRF significantly decreased the concentrations of total bile acid (BA) in the HPV (p < 0.01), JV (p < 0.01), bile (p < 0.05) and SI (p < 0.05), but with no effect on ileal BAs excretion flux. Furthermore, blood triglyceride (TAG) levels were also lower with AXRF (p < 0.01) but with no significant effects on LDL-, HDL- or total cholesterol levels. The lower plasma TAG concentration was consistent with the reduced/delayed digestion and absorption of TAG with the AXRF (total fatty acid and MUFA p < 0.01; unsaturated fatty acid p < 0.05). Conclusion: The results suggest that AXRF reduced the levels of circulating BAs which slowed down the digestion of TAG and absorption of free fatty acids, with consequent reduction in blood TAG. Reduction in circulating bile acids by arabinoxylan causes reduction in lipids digestion and absorption. Wheat arabinoxylan in the diet masks the presence of lipids in the small intestine. This results in a decrease in bile acids secretion by the gall bladder after an adaption period. The rate of lipids digestion and absorption is thus reduced with a subsequent reduction in blood trigycerides.

The human gut contains over 100 trillion bacteria comprising over 1000 bacterial species and in excess of 1 million genes. Recent research suggests that changes in the gut microbial population structure are associated with a wide range of human diseases including both diseases of the gut, such as colorectal cancer, irritable bowel syndrome and inflammatory bowel diseases, and systemic disorders such as obesity, diabetes, cardiovascular disease, autism and Parkinson’s disease. One of the most potent modifiers of gut microbiota structure and function is food. Using dietary fibre as a paradigm, we discuss how foods can modify key health-related functions through their interaction with the gut microbiota and resultant metabolites that are formed.

The relationships between starch structure and functionality are important in underpinning the industrial and nutritional utilisation of starches. In this work, the relationships between the biosynthesis, structure, molecular organisation and functionality have been examined using a series of defined genotypes in barley with low (20%), standard (20-30%), elevated (30-50%) and high (50%) amylose starches. A range of techniques have been employed to determine starch physical features, higher order structure and functionality. The two genetic mechanisms for generating high amylose contents (down-regulation of branching enzymes and starch synthases, respectively) yielded starches with very different amylopectin structures but similar gelatinisation and viscosity properties driven by reduced granular order and increased amylose content. Principal components analysis (PCA) was used to elucidate the relationships between genotypes and starch molecular structure and functionality. Parameters associated with granule order (PC1) accounted for a large percentage of the variance (57%) and were closely related to amylose content. Parameters associated with amylopectin fine structure accounted for 18% of the variance but were less closely aligned to functionality parameters.

Animal studies show that increasing large bowel butyrate concentration through ingestion of butyrylated or resistant starches opposes carcinogen-induced tumorigenesis, which is consistent with population data linking greater fiber consumption with lowered colorectal cancer (CRC) risk. Butyrate has been shown to regulate the apoptotic response to DNA damage. This study examined the impact of increasing large bowel butyrate concentration by dietary butyrylated starch on the colonic epithelium of rats treated with the genotoxic carcinogen azoxymethane (AOM). Four groups of 10 male rats were fed AIN-93G based-diets containing either low amylose maize starch (LAMS), LAMS with 3% tributyrin, 10% high amylose maize starch (HAMS) or 10% butyrylated HAMS (HAMSB). HAMS and HAMSB starches were cooked by heating in water. After 4 weeks, rats were injected once with AOM and killed 6 h later. Rates of apoptosis and proliferation were measured in colonic epithelium. Short-chain fatty acid concentrations in large bowel digesta and hepatic portal venous plasma were higher in HAMSB than all other groups. Apoptotic rates in the distal colon were increased by HAMSB and correlated with luminal butyrate concentrations but cellular proliferation rates were unaffected by diet. The increase in apoptosis was most marked in the base and proliferative zone of the crypt. Regulation of luminal butyrate using HAMSB increases the rates of apoptotic deletion of DNA-damaged colonocytes. We propose this pro-apoptotic function of butyrate plays a major role reducing tumour formation in the AOM-treated rat and that these data support a potential protective role of butyrate in CRC.

BACKGROUND Short-chain fatty acids (SCFAs) maintain human colonic function and may help prevent colonic disease. A study with ileostomists showed that starches acylated with specific SCFAs largely survive passage through the small intestine, but the percentage released in the colon has not been established. OBJECTIVE The objective was to determine the percentage of ingested esterified butyrate released in the human gastrointestinal tract. DESIGN The study was a randomized, crossover, controlled trial consisting of baseline and four 2-wk periods during which 16 volunteers consumed diets low in resistant starch plus 20 and 40 g cooked high-amylose maize starch (HAMS: HAMS20 or HAMS40) or butyrylated HAMS (HAMSB20 or HAMSB40) daily. HAMSB20 contained 31.8 mmol esterified butyrate. Complete 48-h fecal collections were made on days 2-3 and 12-13 of each period. RESULTS Free fecal butyrate concentrations were higher after HAMSB40 than after HAMSB20 (P < 0.005) and HAMS (P < 0.0001) and higher than baseline data (P < 0.0001). Fecal esterified butyrate concentrations were highest in the HAMSB40 (days 12-13; P < 0.0001) group, and concentrations in the HAMSB40 (days 2-3) and HAMSB20 groups were higher than those in the HAMS groups and those at baseline (P < 0.0001). Ingestion of HAMSB20 and HAMSB40 resulted in the release of 26.8 ± 1.0 and 50.2 ± 2.4 mmol butyrate/d (days 12-13) (84.2 ± 3.0% and 79.0 ± 3.1% of total ingested esterified butyrate), respectively, in the gastrointestinal tract. By calculation, ∼57.2% of ingested esterified butyrate was released in the colon. Microbial analysis showed that this release was probably facilitated mainly by Parabacteroides distasonis, which increased in abundance with HAMSB40 (days 12-13) (P < 0.001). CONCLUSIONS This study shows that cooked butyrylated starch delivers esterified butyrate to the human colon effectively and has the potential to improve human bowel health. This trial is registered in the Australian Clinical Trials Registry as ACTRN012606000398505.

The inactivation of starch branching IIb (SBEIIb) in rice is traditionally associated with elevated apparent amylose content, increased peak gelatinization temperature, and a decreased proportion of short amylopectin branches. To elucidate further the structural and functional role of this enzyme, the phenotypic effects of down-regulating SBEIIb expression in rice endosperm were characterized by artificial microRNA (amiRNA) and hairpin RNA (hp-RNA) gene silencing. The results showed that RNA silencing of SBEIIb expression in rice grains did not affect the expression of other major isoforms of starch branching enzymes or starch synthases. Structural analyses of debranched starch showed that the doubling of apparent amylose content was not due to an increase in the relative proportion of amylose chains but instead was due to significantly elevated levels of long amylopectin and intermediate chains. Rices altered by the amiRNA technique produced a more extreme starch phenotype than those modified using the hp-RNA technique, with a greater increase in the proportion of long amylopectin and intermediate chains. The more pronounced starch structural modifications produced in the amiRNA lines led to more severe alterations in starch granule morphology and crystallinity as well as digestibility of freshly cooked grains. The potential role of attenuating SBEIIb expression in generating starch with elevated levels of resistant starch and lower glycaemic index is discussed.

Butyrate and other SCFA produced by bacterial fermentation of resistant starch (RS) or nonstarch polysaccharides (NSP) promote human colonic health. To examine variation in fecal variables, especially butyrate, among individuals and the response to these fibers, a randomized cross-over study was conducted that compared the effects of foods supplying 25 g of NSP or 25 g of NSP plus 22 g of RS/d over 4 wk in 46 healthy adults (16 males, 30 females; age 31-66 y). Fecal SCFA levels varied widely among participants at entry (butyrate concentrations: 3.5-32.6 mmol/kg; butyrate excretions: 0.3-18.2 mmol/48 h). BMI explained 27% of inter-individual butyrate variation, whereas protein, starch, carbohydrate, fiber, and fat intake explained up to 16, 6, 2, 4, and 2% of butyrate variation, respectively. Overall, acetate, butyrate, and total SCFA concentrations were higher when participants consumed RS compared with entry and NSP diets, but individual responses varied. Individual and total fecal SCFA excretion, weight, and moisture were higher than those for habitual diets when either fiber diet was consumed. SCFA concentrations (except butyrate) and excretions were higher for males than for females. Butyrate levels increased in response to RS in most individuals but often decreased when entry levels were high. Fecal butyrate and ammonia excretions were positively associated ((2) = 0.76; P < 0.001). In conclusion, fecal butyrate levels vary widely among individuals but consuming a diet high in RS usually increases levels and may help maintain colorectal health.

It is well established that inulin-type fructans, such as oligofructose derived from chicory root, improve gastrointestinal health via fermentation and proliferation of desirable bacteria in the large bowel. However, differences in fructan structure may contribute significantly to different fermentation and subsequent metabolic effects. Here we ascertain the chain length of two novel cereal fructans (graminan) isolated from wheat stem and barley grain, and investigate their fermentation capacity in vitro , in comparison to inulin and oligofructose. Fractions, representing three predominant fructo-oligosaccharide (FOS) size ranges from each source, were fermented and short chain fatty acids (SCFA) production was measured after 24 h of batch culture. Graminan FOS when fermented produced similar levels of total SCFA as oligofructose and inulin. The fermentation of high DP FOS fractions of barley grain and wheat stem (DP 5–15) produced more propionate than oligofructose. Graminan fructans may provide a useful alternative source of oligofructose with potential additional health benefits.

Cell walls in commercially important cereals and grasses are characterized by the presence of (1,3;1,4)-β-d-glucans. These polysaccharides are beneficial constituents of human diets, where they can reduce the risk of hypercholesterolemia, type II diabetes, obesity and colorectal cancer. The biosynthesis of cell wall (1,3;1,4)-β-d-glucans in the Poaceae is mediated, in part at least, by the cellulose synthase-like CslF family of genes. Over-expression of the barley CslF6 gene under the control of an endosperm-specific oat globulin promoter results in increases of more than 80% in (1,3;1,4)-β-d-glucan content in grain of transgenic barley. Analyses of (1,3;1,4)-β-d-glucan fine structure indicate that individual CslF enzymes might direct the synthesis of (1,3;1,4)-β-d-glucans with different structures. When expression of the CslF6 transgene is driven by the Pro35S promoter, the transgenic lines have up to sixfold higher levels of (1,3;1,4)-β-d-glucan in leaves, but similar levels as controls in the grain. Some transgenic lines of Pro35S:CslF4 also show increased levels of (1,3;1,4)-β-d-glucans in grain, but not in leaves. Thus, the effects of CslF genes on (1,3;1,4)-β-d-glucan levels are dependent not only on the promoter used, but also on the specific member of the CslF gene family that is inserted into the transgenic barley lines. Altering (1,3;1,4)-β-d-glucan levels in grain and vegetative tissues will have potential applications in human health, where (1,3;1,4)-β-d-glucans contribute to dietary fibre, and in tailoring the composition of biomass cell walls for the production of bioethanol from cereal crop residues and grasses.

Dietary red and processed meats may increase risk of colorectal cancer (CRC), whereas fiber may be protective. Recently, we demonstrated that dietary beef causes greater colonic DNA strand breakage than equivalent levels of chicken in rats and that resistant starch (RS) as 20% high amylose maize starch (HAMS) attenuated the damage. From that study, we now report measures of circulating factors that may influence CRC initiation or progression including malondialdehyde (MDA), leptin, insulin-like growth factor-I (IGF-I), insulin, matrix metalloproteinase-2 (MMP-2), tissue inhibitor of MMP-2 (TIMP-2), interleukins (IL), and short chain fatty acids. MDA levels were increased by beef diets relative to the chicken diets. Leptin concentrations, which were lower for chicken than beef at the 35% level in the absence of HAMS, were lowered by HAMS. Higher dietary chicken (but not beef) increased IGF-I irrespective of HAMS feeding. Higher levels of chicken resulted in greater insulin concentrations than for beef in rats fed HAMS. Without dietary HAMS, TIMP-2 concentration increased in response to both meats but was highest for chicken. MMP-2 and TIMP-2 concentrations were higher for HAMS diets. IL-1beta and IL-12 concentrations were lowered by HAMS feeding. Colonic DNA strand breakage was positively associated with circulating leptin and MDA concentrations as well as tissue MDA concentrations and negatively associated with plasma TIMP-2 concentration. MMP-2 and TIMP-2 positively correlated with hepatic portal butyrate levels but leptin concentrations correlated negatively. These results suggest diets high in meat or RS could influence cancer initiation or progression by changes in circulating levels of hormones and other factors.

Factors controlling the concentration of resistant starch (RS) in foods are of considerable interest on account of the potential for this type of fibre to deliver health benefits to consumers. The present study was aimed at establishing changes in starch granule morphology as a result of human small-intestinal digestion. Volunteers with ileostomy consumed six selected foods: breakfast cereal (muesli), white bread, oven-baked French fries, canned mixed beans and a custard containing either a low-amylose maize starch (LAMS) or a high-amylose maize starch (HAMS). Analysis showed that digesta total RS (as a fraction of ingested starch) was: muesli, 8·9 %; bread, 4·8 %; fries, 4·2 %; bean mix, 35·9 %; LAMS custard, 4·0 %; HAMS custard, 29·1 %. Chromatographic analysis showed that undigested food contained three major starch fractions. These had average molecular weights (MW) of 43 500 kDa, 420 kDa and 8·5 kDa and were rich in amylopectin, higher-MW amylose and low-MW amylose, respectively. The low-MW amylose fraction became enriched preferentially in the stomal effluent while the medium-MW starch fraction showed the greatest loss. Fourier transform IR spectroscopy showed that absorbance at 1022 per cm decreased after digestion while the absorbance band at 1047 per cm became greater. Such changes have been suggested to indicate shifts from less ordered to more ordered granule structures. Further analysis of amylose composition by scanning iodine spectra indicated that the MW of amylose in ileal digesta was lower than that of undigested amylose. It appears that high-MW amylose is preferentially digested and that MW, rather than amylose content alone, is associated with resistance of starch to digestion in the upper gut of humans.

Fresh and stored melons were analyzed for total and bioaccessible beta-carotene content, as well as total antioxidant potentials, to investigate the effects of genotype, season, sowing time, and storage on nutritional quality. There were significant cultivar effects on beta-carotene content (P0.001), ranging from 1 to 23 mg kg(-1) of fresh weight (FW), its bioaccessibility (P0.005), ranging from 79 to 94.5%, and for antioxidant potential (P0.001), ranging from 1569 to 2267 micromol of Fe(2+) kg(-1) of FW of melons. Sowing time significantly affected beta-carotene contents (P0.017) and antioxidant potentials (P0.001), but these were highly cultivar dependent, with season having no effect. Postharvest storage over 4 weeks at 7 degrees C resulted in losses of antioxidant potential and beta-carotene content independent of cultivar. Cultivar and postharvest management were major determinants of nutritional quality of the melons evaluated.

In this article, we describe a simple, sensitive, accurate, and repeatable method for the measurement of phenol and p-cresol (4-methylphenol) in human urine and feces. We examined a number of parameters to identify an optimal extraction protocol. Purification of sample extracts was achieved by low-temperature vacuum microdistillation. Separation was achieved in approximately 15 min by high-performance liquid chromatography (HPLC) with quantification by fluorescence at 284/310 nm. Limits of detection for phenol were 2 ng/ml for urine and 20 ng/g for feces, and those for p-cresol were 10 ng/ml for urine and 100 ng/g for feces. For comparison, approximate mean values for urine are 3 microg/ml for phenol and 30 microg/ml for p-cresol, and those for feces are 1 microg/g for phenol and 50 microg/g for p-cresol. An experienced analyst can process 60 samples each day using this method.

The amylase digestibility of high-amylose maize starches has been compared before and after thermomechanical processing. Starches were analysed for enzyme-resistant starch yield, apparent amylose content, crystallinity (X-ray diffraction), and molecular order (NMR and FTIR), both before and after treatment with a-amylase. All samples had significant (>10%) enzyme-resistant starch levels irrespective of the type and extent of thermal or enzymic processing. Molecular or crystalline order was not a pre-requisite for enzyme resistance. Near-amorphous forms of high amylose maize starches are likely to undergo recrystallisation during the enzyme-digestion process. The mechanism of enzyme resistance of granular high-amylose starches is found to be qualitatively different to that for processed high-amylose starches. For all samples, measured levels of enzyme resistance are due to the interruption of a slow digestion process, rather than the presence of completely indigestible material. 2008 Elsevier Ltd. All rights reserved.

Pigs were fed a freeze-dried probiotic (Bifidobacterium animalis CSCC 1941) plus a high-amylose maize starch (HAMS) and a fructooligosaccharide (FOS) separately or together. Fecal output and total and individual major short-chain fatty acid (SCFA) concentrations and excretion were higher and pH was lower with HAMS than with FOS relative to when they were fed a low-amylose maize starch (LAMS; control). Fecal bifidobacteria numbers and total excretion were equally higher during feeding of FOS or HAMS and highest with HAMS + FOS. When probiotic supplementation was stopped, bifidobacteria numbers declined rapidly when they were fed LAMS, more slowly with FOS or HAMS, and were maintained with HAMS + FOS. The data confirm that both HAMS and FOS are prebiotics and suggest that they act through different mechanisms and that they are most effective in combination. However only HAMS raises fecal SCFA.

Summary Numerous in vitro carbohydrate digestion methods exist for analysis of the likely glycaemic properties of foods. Generally these methods encompass simulations of oral, gastric and intestinal digestion processes, but the way in which physiological conditions are implemented across methods differs considerably. Some differences are in the mode of comminution, inclusion and duration of a gastric digestion, and choice of amylolytic enzyme. Incubation temperature, pH, duration and stirring mode also differ between methods. Such differences, particularly the method used to mimic chewing, can have a substantial influence on the relative estimate of glycaemic potency for a given food. To achieve estimates of high predictive power, and global relevance, a validated, standardised in vitro digestion method must be developed. This article provides a systematic review of commonly used and referred to in vitro carbohydrate digestion methods. Methodological discrepancies between protocols are identified thus defining the route a systematic standardisation investigation should take.

Himalaya 292 is a novel hull-less barley cultivar with lower total starch content, a higher proportion of amylose and a substantially higher content of soluble and insoluble fibre. The high fibre content suggests that the grain may have potential as an ingredient in health promoting foods. In this study, 29 healthy volunteers consumed extruded whole grain breakfast cereal made from Himalaya 292 (123 g) or a commercial hull-less barley (cv Torrens) (76 g) each supplying 50 g carbohydrate. Glycemic indices were 77 ± 13 (SEM) and 50 ± 9 for Torrens and Himalaya 292 breakfast cereals respectively (P = 0.004). On this basis, the Himalaya 292 cereal is a low glycemic index food. The glycemic load (the product of glycemic index and the available carbohydrate content of a 60 g serve divided by 100) of the new cultivar was 10.1 ± 1.8 compared to 25.3 ± 4.3 for the Torrens breakfast cereal (P < 0.001). The plasma insulin response to the test meal was 26% lower for Himalaya 292 than for Torrens (P = 0.023). The data confirm that Himalaya 292 may be of value in foods designed to assist in the prevention and management of diabetes.

Colorectal cancer (CRC) is a major cause of death worldwide. Studies suggest that dietary fibre offers protection perhaps by increasing colonic fermentative production of butyrate. This study examined the importance of butyrate by investigating the effects of resistant starch (RS) and butyrylated-RS on azoxymethane (AOM)-induced CRC in rats. Four groups (n = 30 per group) of Sprague–Dawley rats were fed AIN-93G-based diets containing a standard low-RS maize starch (LAMS), LAMS + 3% tributyrin (LAMST), 10% high-amylose maize starch (HAMS) and 10% butyrylated HAMS (HAMSB) for 4 weeks. Rats were injected once weekly for 2 weeks with 15 mg/kg AOM, maintained on diets for 25 weeks and then killed. Butyrate concentrations in large bowel digesta were higher in rats fed HAMSB than other groups (P < 0.001); levels were similar in HAMS, LAMS and LAMST groups. The proportion of rats developing tumours were lower in HAMS and HAMSB than LAMS (P

High pressure processing (HPP) is a relatively new food preservation processing technology that enhances food safety and shelf-life without compromising organoleptic qualities. There has been little research on the impact of HPP on the nutritional and health-promoting properties of foods to date and most of it has focused on juices and purees of fruit such as oranges and tomatoes. The objective of this study was to determine the effects of HPP treatment at two pressure levels (400 MPa; 600 MPa) on antioxidant activity, total carotenoid content and carotenoid availability in vitro, of three commonly consumed vegetables. Antioxidant capacity and total carotenoid content differed between vegetables but were unaffected by HPP treatment. In vitro availability of specific carotenoids also varied greatly between vegetables (3–35%). HPP altered availability of carotenoids according to the type of vegetable treated and processing pressure applied, however the magnitude of the responses was minor. Industrial relevance This study provides further scientific evidence of the benefits of high pressure processing in retaining the nutritional attributes of fresh foods. Antioxidant activity and levels of carotenoids before and after exposure to high pressures (up to 600 MPa for 2 min) were essentially no different. Also, the data suggest that micronutrients and phytochemicals in certain vegetables may be made more bioavailable by high pressure treatment. From a nutritional perspective, high pressure processing is an attractive food preservation technology and clearly offers opportunities for horticultural and food processing industries to meet the growing demand from consumers for healthier food products.

Different types of novel wheat lines with different starch contents and amylose/amylopectin ratios, relating to defined alterations in the number and activity of starch synthase IIa genes, were processed by pilot-plant extrusion. Two types of products were produced: pure wholemeal products and breakfast cereals made from wholemeal/maize blends. Lower apparent shear viscosity was obtained in the extruder with lower starch content and higher amylose/amylopectin ratio flours (SSIIa-deficient line). The bulk density of the products decreased with increasing extrusion temperature and was always higher for the triple-null line. The bulk density was not completely explained by the melt shear viscosity, suggesting the importance of the fillers (fibers, brans) in the process of expansion and structure acquisition. The different mechanical properties were explained by the density and by the material constituting the cell walls. Enzyme-resistant starch (RS) content and hydrolysis index (HI) were not correlated to the extrusion temperature, but RS was higher in pure wholemeal products and in the SSIIa-deficient line. These results are discussed in terms of starch molecular architecture and product microstructure.

Some of the starch consumed by humans is not digested in the small intestine. Such starch, known as resistant starch, is fermented in the large intestine and leads to the production of short chain fatty acids. Increased consumption of resistant starch is associated with improved cardio-vascular health. A high proportion of amylose in the starch consumed is correlated with increased resistant starch but other unknown aspects of starch structure may also influence the digestibility of starch. Detailed investigation of the starch biosynthetic pathway has revealed that reducing the activity of specific isoforms of branching enzymes and starch synthases can lead to increased amylose. Methods to alter the expression of and detect mutations in targeted genes involved are discussed.

Human population studies show that dietary red and processed, but not white, meats are associated with increased risk of colorectal cancer but dietary fibre appears to be protective. We examined whether dietary cooked red or white meat had differential effects on colonic DNA damage in rats and if resistant starch (RS), a dietary fibre component, provided protection. Rats were fed diets containing approximately 15, 25 or 35% of cooked beef or chicken, both with or without 20% high-amylose maize starch (HAMS) as a source of RS, for 4 weeks. DNA single-strand breaks (SSB) and double-strand breaks (DSB) were measured in isolated colonocytes (by comet assay) along with apoptosis levels, colonic mucus thickness and large bowel short-chain fatty acids (SCFA). Both red and white meat increased colonocyte SSB and DSB dose dependently but damage was substantially greater with red meat. Dietary HAMS prevented these increases. Apoptotic cell numbers were increased dose dependently by red meat irrespective of HAMS feeding, whereas white meat only increased apoptotic cell numbers in the presence of HAMS. Red meat induced greater colonic mucus layer thinning than white meat but HAMS was protective in both cases. HAMS induced increases in large bowel SCFA, including butyrate, and significantly lowered concentrations of phenols and cresols. We have demonstrated that dietary red meat causes greater levels of colonic DNA SSB and DSB than white meat, consistent with the epidemiological data. Dietary RS protects against this damage and also against loss of the mucus barrier, probably through increased butyrate production.